Difference between revisions of "C2AB: A Molecular Glue for Lipid Vesicles with a Negatively Charged Surface"

From Soft-Matter
Jump to: navigation, search
(Soft matter discussion)
(Soft matter discussion)
Line 19: Line 19:
 
The academic group conducting this research is experimenting with liposomes as a drug delivery vehicle.  They believe they can solve the problem of targeting the liposome to the correct cells, however there is a perceived problem that the concentration of the drug in each liposome will not have the necessary efficacy.  They hope to be able to fuse the liposomes together at the site of the target to ensure that the drug concentration is high enough to provide a sufficient dosage.
 
The academic group conducting this research is experimenting with liposomes as a drug delivery vehicle.  They believe they can solve the problem of targeting the liposome to the correct cells, however there is a perceived problem that the concentration of the drug in each liposome will not have the necessary efficacy.  They hope to be able to fuse the liposomes together at the site of the target to ensure that the drug concentration is high enough to provide a sufficient dosage.
  
They are collaborating this work with a group that has been studying the protein C2AB which is used in cells as a calcium sensor in synaptic vesicle exocytosis.  The protein has two active sites, one of which binds 2 <math>Ca^{2+}</math>, the other binding 3 <math>Ca^{2+}</math>.
+
They are collaborating this work with a group that has been studying the protein C2AB which is used in cells as a calcium sensor in synaptic vesicle exocytosis.  The protein has two active sites, one of which binds 2 <math>Ca^{2+}</math>, the other binding 3 <math>Ca^{2+}</math>. When this protein binds the <math>Ca^{2+}</math> it undergoes a conformational change that creates two positively charged ends that are very strongly attracted to negatively charged liposomes.
 +
 
 +
The group created negatively surface charged liposomes and anchored them to a quartz substrate.  They then fluorescently labeled these liposomes.  After this they fluorescently labeled other liposomes with a molecule emitting a different wavelength.  When they put these two liposomes together, they did not bind to eachother as determined by fluorimetry.  After adding C2AB to the mixture, in the absence of <math>Ca^{2+}</math> still no binding occurred.  However, when <math>Ca^{2+}</math> was added, binding took place immediately, within two seconds.  They then used <math>Mg^{2+}</math> in place of <math>Ca^{2+}</math> to determine if it was the existence of any divalent cation that caused the binding.  The <math>Mg^{2+}</math> however did not cause binding.  This shows that the C2AB is only activated as a 'glue' in the presence of <math>Ca^{2+}</math>, and it is the positive charge of the <math>Ca^{2+}</math> along with the conformational change of the C2AB that is causing this activation.

Revision as of 01:42, 3 November 2009

Original entry: William Bonificio, AP 225, Fall 2009

Information

C2AB: A Molecular Glue for Lipid Vesicles with a Negatively Charged Surface. Jiajie Diao, Tae-Young Yoon, Zengliu Su, Yeon-Kyun Shin, Taekjip Ha. Langmuir 2009 25 (13), 7177-7180

Soft matter keywords

Liquid-Phase Crystallization, Pentazene, Organic Field Effect Transistor, Pentacene, Tricholorbenzene.

Summary

The purpose of this study was to test the efficacy of a calcium binding protein, currently found in cells, to fuse liposomes to one another. The binding protein, C2AB, binds calcium at both ends of it. This binding gives it a positive charge on both sides, which in turn gives the C2AB an affinity to negatively charged liposomes. When liposomes, C2AB and <math>Ca^{2+}</math> are combined the liposomes fuse.

Soft matter discussion

Schematic illustration of experiment. Immobilized vesicles 'glued' to mobile vesicles via C2AB.


(a) shows TIR images before and after the introduction of <math>Ca^{2+}</math>. (b) and (c) show TEM images of the liposomes before and after the addition of <math>Ca^{2+}</math>

The academic group conducting this research is experimenting with liposomes as a drug delivery vehicle. They believe they can solve the problem of targeting the liposome to the correct cells, however there is a perceived problem that the concentration of the drug in each liposome will not have the necessary efficacy. They hope to be able to fuse the liposomes together at the site of the target to ensure that the drug concentration is high enough to provide a sufficient dosage.

They are collaborating this work with a group that has been studying the protein C2AB which is used in cells as a calcium sensor in synaptic vesicle exocytosis. The protein has two active sites, one of which binds 2 <math>Ca^{2+}</math>, the other binding 3 <math>Ca^{2+}</math>. When this protein binds the <math>Ca^{2+}</math> it undergoes a conformational change that creates two positively charged ends that are very strongly attracted to negatively charged liposomes.

The group created negatively surface charged liposomes and anchored them to a quartz substrate. They then fluorescently labeled these liposomes. After this they fluorescently labeled other liposomes with a molecule emitting a different wavelength. When they put these two liposomes together, they did not bind to eachother as determined by fluorimetry. After adding C2AB to the mixture, in the absence of <math>Ca^{2+}</math> still no binding occurred. However, when <math>Ca^{2+}</math> was added, binding took place immediately, within two seconds. They then used <math>Mg^{2+}</math> in place of <math>Ca^{2+}</math> to determine if it was the existence of any divalent cation that caused the binding. The <math>Mg^{2+}</math> however did not cause binding. This shows that the C2AB is only activated as a 'glue' in the presence of <math>Ca^{2+}</math>, and it is the positive charge of the <math>Ca^{2+}</math> along with the conformational change of the C2AB that is causing this activation.